Spheroid and Tissue Assembly via Click Chemistry in Microfluidic Flow

Abstract

Proper cell-cell contact and communication are essential for the correct development and survival of higher order organisms. In order to study complex cell interactions that occur in vivo, model systems that are able to recapitulate 3D cell-cell interactions in vitro are key to advancing new biotechnologies, therapeutics, and tissue engineering applications. Herein, we show a new strategy to rapidly and efficiently generate complex multiple cell line containing spheroids and tissues in microfluidic flow without the use of scaffolds, molecular biology, or metabolic biosynthesis. The method relies on the integration of microfluidics, liposome fusion, bio-orthogonal chemistry, and cell surface engineering to rapidly click coculture cell assemblies in flow. We demonstrate this strategy by assembling various combinations of cell types with an interfacial cell to cell click chemistry in microfluidic flow to generate a range of spheroid types and oriented tissue multilayers.

Tumour-on-a-chip: microfluidic models of tumour morphology, growth and microenvironment

“…Similarly, to produce tumour spheroids by avoiding cell adhesion to cultureware and inducing aggregation, an aqueous two-phase system can also compartmentalize cell suspension and produce spheroids without the concern of drying and possible inefficiency in chemical transport and toxicity of an oil phase [51][52][53]. Three-dimensional spheroids can also be formed by assembly of cells using bio-orthogonal chemistry [54] or incubation of cells with magnetic particles [55,56] (figure 2g,h). Recently, several microfluidic techniques have been developed to create tumour spheroids by either hydrodynamic trapping of cells in stagnation regions or in microwell structures [57][58][59][60], aggregating multiple cells in double-emulsions or hydrogel droplets [61][62][63][64], or aggregating cells on a digital microfluidic platform [65] (figure 2i).…”

Scaffold Free Bio-orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering

“…In order to generate scaffold free functional 3-dimensional cardiac tissue, we used the combination of liposome fusion, cell surface engineering and bio-orthogonal chemistry20212223. We have previously shown the rapid installation of bio-orthogonal ketone and oxyamine groups to a range of cell types via liposome fusion (ViaGlue)242526.…”

“…These tissues may be used for many applications including drug screening and as models for disease and infection as well as eventual cardiac patch in vivo applications. Many different cell types including stem cells may be used with the strategy and the inter cell click ligation is compatible with microfluidic and 3D printing technologies2324252627282930. We believe the combination of liposome fusion, bio-orthogonal chemistry and cell surface engineering to tailor cell surfaces will have a significant impact on autocrine and paracrine signaling studies and for the development and evaluation of tissues for drug screening and therapeutic organ on a chip based biotechnology applications4041424344.…”

Programming Cell-Cell Interactions through Non-genetic Membrane Engineering

“…Cells bearing the complementary ketone and oxyamine moieties were then linked to one another via a covalent oxime ligation. When applied to multiple cell types, the Yousaf group has been able to form multicellular sheets (Dutta et al, 2011b), spheroids (O’Brien et al, 2015), and microtissues (Rogozhnikov et al, 2016). …”